缺氧所致海马神经元N-甲基-D-天冬氨酸2B受体和PTEN基因的表达  

Expression of NR2B and PTEN in rat hippocampus neurons subjected to anoxia insult

作　　者：陈力学[1] 刘宝松[2] 周冀英[1] 桂蓓[1] 邵阳[2] 

机构地区：[1]重庆医科大学附属第一医院实验研究中心重庆市神经病学重点实验室,重庆400016 [2]第三军医大学大坪医院野战外科研究所三室,重庆400042

出　　处：《中风与神经疾病杂志》2010年第4期296-298,共3页Journal of Apoplexy and Nervous Diseases

基　　金：重庆市科委自然科学基金资助项目(2007BB5287);重医一院院内医学科学基金项目(YXJJ2009-15)

摘　　要：目的探讨OGD缺氧损伤后大鼠海马神经元NR2B受体和抑癌基因PTEN的表达量,以及NR2B与PTEN是否有生理性相互作用关系,为研究PTEN基因在脑缺血中的保护作用机制奠定基础。方法建立海马神经元体外OGD损伤模型,采用细胞比色分析(colorimetric assay)法和Western blotting检测神经元NR2B含量,RT-PCR和Western blotting测定PTEN基因的mRNA和蛋白表达水平,免疫沉淀和Western blotting研究NR2B与PTEN的相互作用。结果细胞比色分析和Western blotting结果显示神经元有大量NR2B表达,OGD损伤72h后NR2B含量表达量明显减少(P<0.05);Western blotting和RT-PCR结果显示OGD损伤后PTEN表达量略有下降但尚无统计学意义(P>0.05),免疫沉淀显示PTEN与NR2B在物理上连接形成复合物。结论海马神经元膜表面NR2B含量丰富,PTEN基因在海马神经元里有表达,OGD缺氧损伤再灌注引起NR2B受体和PTEN基因表达量下降,PTEN与NR2B有生理性相互作用。Objective To investigate the expression of N-methyl-D-aspartate receptor（NMDA）2B and PTEN on rat hippocampus neurons with OGD model,and confirm whether there is physical association between PTEN and NR2B in rat hippocampus.Methods In the OGD model with anaerobic gas mixture and deoxygenated,glucose-free extracellular solution,the quantities of NR2B in the neuronal membrane were measured by colorimetric assay and western blotting.The expression of PTEN were analyzed by western blotting and RT-PCR.The physical association between PTEN and NMDARs in rat hippocampus was determined by western blotting and immunoprecipitation.Results The study demonstrated that there were numerous NR2B subunit expression on neuronal membrane by OPD assay and Western blotting.There was significantly decrease of the expression and quantity of NR2B at 72 hr after oxygen and glucose deprivation（OGD）and reperfusion（P0.05）.The expression level of PTEN decreased after anoxic injury and reperfusion,but no statistical difference compared with the control group emerged.There were physical association between PTEN and NR2B in rat hippocampal neuron.Conclusion Our study suggests that there be numerous expression of NR2B and PTEN in rat hippocampal neuron,which show decrease after OGD damage,and there be direct protein-protein interaction between PTEN and NR2B.

关 键 词：PTEN基因 NR2B受体 OGD 细胞比色分析 

分 类 号：R741[医药卫生—神经病学与精神病学]